基于“心肾相关”理论的鹿红方通过调控线粒体合成改善心梗后心力衰竭大鼠心肌能量代谢的机制研究

Heart failure is one of the most difficult cardiovascular disease to date.There are obvious energy metabolism disorders in heart failure,and the links in the energy metabolism disorder have become the latest research targets.Previous studies have confirmed that Luhong formula can improve heart failure by activating PI3K/Akt/eNOS pathway and regulating the function of myocardial mitochondria to intervent energy metabolism.Preliminary observation found that Luhong formula can increase hemoglobin(which is regulated by EPO) in patients with heart.failure.Recently,it has been found that when myocardium is during hypoxia,the secretion of erythropoietin(EPO) is increased.After the combination of EPO and EPOR,PI3K is activated and can make phosphorylation of Akt,and they can increase mitochondrial synthesis.The source and function of EPO is associated with kidney essence in Traditional Chinese Medicine theories.And Luhong formula which is based.on the “heart kidney correlation" theory can supplement the kidney,replenish essence and quicken the blood.Since mitochondria is important organelle for energy metabolism,and energy metabolism is the hot spot of current research on heart failure mechanism.The hypothesis is proposed that Luhong formula can significantly improve the energy metabolism disorder in heart failure.The mechanism may regulate.mitochondrial synthesis by increasing EPO、EPOR content and expression to activate PI3K/Akt/eNOS pathway.From the overall level and cell level to demonstrate the effect of Luhong formula interfereing with the pathway,and to provide basis of new target for prevention and treatment of heart failure by traditional Chinese medicine.

心力衰竭是至今未被攻克的心血管疾病。心衰时存在明显的能量代谢障碍，能量代谢障碍中的各环节已成为最新研究靶点。前期研究证实鹿红方能激活PI3K/Akt/eNOS通路、调节心肌线粒体功能干预能量代谢从而改善心衰。前期观察发现鹿红方能增加心衰患者血红蛋白（其生成受EPO调控）水平。近来研究发现，心肌缺氧时促红细胞生成素（EPO）分泌增多，与EPOR结合后激活PI3K使Akt磷酸化，增加线粒体合成。EPO来源和功能与中医藏象理论中肾精具有同一性，而基于“心肾相关”理论的鹿红方具有补肾益精活血功效。鉴于线粒体是重要的能量代谢细胞器，结合“能量代谢”这一目前研究心衰机制的热点，提出“鹿红方能显著改善心衰时能量代谢障碍，其机制可能是通过增加EPO、EPOR含量和表达，激活PI3K/Akt/eNOS通路调控线粒体合成”假说。拟从整体和细胞水平论证鹿红方对该途径的干预作用，为中医药防治心衰新靶点提供依据。

心力衰竭是至今未被攻克的心血管疾病。现代研究认为，心力衰竭是一种慢性代谢病。心力衰竭时存在底物利用障碍和能量物质缺乏，随之出现包括高能磷酸盐含量、线粒体功能改变在内的心脏能量转换紊乱，而线粒体的功能是整个代谢环节的关键。因此，明确心力衰竭时心肌能量代谢的机制将成为心力衰竭治疗的新靶点。心肌缺氧时促红细胞生成素（EPO）分泌增多，与其靶细胞膜表面的EPOR结合后激活PI3K使Akt磷酸化，增加线粒体合成。EPO来源和功能与中医藏象理论中肾精具有同一性，而基于“心肾相关”理论的鹿红方具有补肾益精活血功效。本研究通过在体实验（心梗后心衰模型）和离体实验（心肌细胞培养），从整体和细胞水平研究鹿红方通过增加EPO、EPOR含量和表达，激活PI3K/Akt/eNOS通路，对线粒体的合成进行调控，进而改善心肌能量代谢，最终改善心力衰竭。第一部分为鹿红方改善心力衰竭心肌能量代谢的体内实验，采取结扎大鼠冠状动脉前降支制备心肌梗死后心力衰竭大鼠模型，造模成功后立即予药物干预，8周后观察鹿红方对EPO生成的影响，剖析EPO对调控线粒体合成的作用机制，揭示鹿红方改善心衰时心肌能量代谢障碍的药效机制。第二部分为鹿红方改善心力衰竭心肌能量代谢的体外实验，制备鹿红方含药血清干预原代心肌细胞，之后再加入一定浓度的EPO干预。观察鹿红方对心肌细胞EPOR表达的影响；剖析EPO对PI3K/Akt/eNOS通路及线粒体合成的调控机制；揭示鹿红方改善心衰时心肌能量代谢障碍的作用机制。通过上述两部分实验，我们验证了“基于‘心肾相关’理论，具有温肾、益精、活血功效的鹿红方可通过增加EPO、EPOR含量和表达，激活PI3K/Akt/eNOS通路调控线粒体的合成，从而改善心力衰竭时心肌的能量代谢。”假说成立，从整体和细胞水平验证了鹿红方改善心力衰竭时心肌能量代谢障碍的机制。从新的视角完善鹿红方对心衰治疗作用的机制研究，同时也为中药治疗能量代谢障碍的机制研究提供新的研究思路。